Department of Chemistry, Yonsei University, Seoul, Korea.
Department of Clinical Pharmacology and Therapeutics, College of Medicine, Kyung Hee University, Seoul, Korea.
Nucleic Acids Res. 2022 Dec 9;50(22):12621-12635. doi: 10.1093/nar/gkac665.
The development of single-cell RNA-seq has broadened the spectrum for biological research by providing a high-resolution analysis of cellular heterogeneity. However, the requirement for sophisticated devices for the compartmentalization of cells has limited its widespread applicability. Here, we develop Onepot-Seq, a device-free method, that harnesses the transient localization of mRNA after lysis to capture single-cell transcriptomes simultaneously in a continuous fluid medium. In mixed-species experiments, we obtained high-quality single-cell profiles. Further, cell type-specific poly(A)-conjugated antibodies allow Onepot-Seq to effectively capture target cells in complex populations. Chemical perturbations to cells can be profiled by Onepot-Seq at single-cell resolution. Onepot-Seq should allow routine transcriptional profiling at single-cell resolution, accelerating clinical and scientific discoveries in many fields of science.
单细胞 RNA 测序的发展通过对细胞异质性进行高分辨率分析，拓宽了生物学研究的范围。然而，细胞分区的复杂设备需求限制了其广泛适用性。在这里，我们开发了一种无设备的方法，即 Onepot-Seq，它利用细胞裂解后 mRNA 的瞬时定位，在连续的流体介质中同时捕获单细胞转录组。在混合物种实验中，我们获得了高质量的单细胞图谱。此外，细胞类型特异性的 poly(A)缀合抗体允许 Onepot-Seq 有效地在复杂群体中捕获靶细胞。通过 Onepot-Seq 可以在单细胞分辨率下对细胞的化学干扰进行分析。Onepot-Seq 应该可以在单细胞分辨率下进行常规转录组分析，从而加速许多科学领域的临床和科学发现。
